布比卡因增强小鼠成肌细胞溶酶体的活性

Bupivacaine can enhance lysosomal activity in mouse muscle myoblasts

目的探讨局部麻醉药布比卡因对小鼠成肌细胞溶酶体的影响。方法将体外培养的小鼠成肌细胞C2C12分为2组。对照组:不加任何药物;布比卡因组:以600μmol/L布比卡因刺激细胞6 h。实验结束后,用Lyso Sensor探针评价溶酶体腔p H,用Lyso Trackor探针检测溶酶体含量,用蛋白免疫印迹法检测溶酶体相关膜蛋白-1(LAMP-1)和溶酶体蛋白水解酶Cathepsin B的表达水平,并以Magic Red染色法测定Cathepsin B的活性。结果布比卡因对溶酶体腔p H没有影响。但是,与对照组相比,布比卡因组溶酶体含量增加15.15%(P<0.01),LAMP-1与Cathepsin B表达量分别增加36.41%、35.29%(P<0.01),Cathepsin B活性增加23.74%(P<0.01)。结论布比卡因能增加小鼠成肌细胞溶酶体含量,增强溶酶体活性。

Objective To investigate the effects of bupivacaine on lysosomal abundance and activity in mouse muscle myoblasts. Methods Mouse myoblasts C2C12 was randomly divided into control group（ without any treatment） and bupivacaine group（ treated with bupivacaine 600 μmol / L for 6 h）. After then,the changes of lysosomal p H was assessed by Lyso Sensor p H indicator. The content of lysosomes was detected by Lyso Tracker probe. The expression of lysosomal-associated membrane protein-1（ LAMP-1） and Cathepsin B was detected by Western blot analysis. The activity of lysosomal proteolytic enzymes Cathepsin B was determined by Magic Red assay kit. Results Bupivacaine did not affect lysosomal p H. However,compared with the controls,lysosomal abundance was significantly increased 15. 15% following bupivacaine treatment（ P〈0. 01）. Moreover,protein expression levels of LAMP-1 and Cathepsin B were significantly upregulated36. 41% and 35. 29% respetctively by bupivacaine（ P〈0. 01）. Furthermore,the activity of Cathepsin B was significantly increased 23. 74% by bupivacaine（ P〈0. 01）. Conclusions Bupivacaine increased lysosomal content and enhance lysosomal activity in mouse muscle myoblasts.